Extended Data Fig. 3: Subtomogram averages and structural comparison of the mES and neural progenitor NPCs.
From: Nuclear pores safeguard the integrity of the nuclear envelope
(a) The schematic data processing workflow for subtomogram averaging of the wild-type mES NPCs. The eight-fold symmetric NPCs from the three other datasets were processed following a similar workflow. (b - e) Composite maps of the eight-fold symmetric NPCs (left) and corresponding Fourier shell correlation curves of the CR, IR and NR averages (right). The data for the wild-type mES NPC (b), the wild-type neural progenitor NPC (c), the Nup133-/- mES NPC (d), and the Nup133-/- neural progenitor NPC (e) are shown. Composite maps are shown as cutaway views. (f, g) Subtomogram-based measurement of the angles between the outer and inner nuclear membranes. In (f), the workflow of measurement is shown as a schematic representation. Based on the subtomogram averages of individual rings (CR, IR, and NR), five coordinates were derived for each NPC subunit (membrane anchor sites of the outer Y-complex of CR and NR (two each from two adjacent protomers) and center of the membrane attachment interface of IR, depicted as green spheres). Subsequently, the angle between two planes (one formed between IR and two CR coordinates, the other formed between IR and two NR coordinates) were measured for each NPC subunit. The measured angles were averaged per NPC, and the averaged angles were plotted (g). NPCs with less than four measured values were discarded from the analysis. Bars denote the means and standard deviations (n = 184 NPCs for the wild-type mES dataset, n = 119 NPCs for the wild-type neural progenitor dataset, n = 128 NPCs for the Nup133-/- mES dataset, n = 136 NPCs for the Nup133-/- neural progenitor dataset). Kruskal-Wallis rank sum test followed by Dunn’s multiple comparisons test. Source numerical data are available in source data.
